CN107446035A - New protein material - Google Patents
New protein material Download PDFInfo
- Publication number
- CN107446035A CN107446035A CN201710552855.9A CN201710552855A CN107446035A CN 107446035 A CN107446035 A CN 107446035A CN 201710552855 A CN201710552855 A CN 201710552855A CN 107446035 A CN107446035 A CN 107446035A
- Authority
- CN
- China
- Prior art keywords
- angiogenin
- hydrolysate
- cystatin
- protein material
- bone
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000000463 material Substances 0.000 title claims abstract description 109
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 101
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 101
- 102100022987 Angiogenin Human genes 0.000 claims abstract description 117
- 108010072788 angiogenin Proteins 0.000 claims abstract description 117
- 239000000413 hydrolysate Substances 0.000 claims abstract description 91
- 102000015833 Cystatin Human genes 0.000 claims abstract description 75
- 108050004038 cystatin Proteins 0.000 claims abstract description 75
- 108010009736 Protein Hydrolysates Proteins 0.000 claims abstract description 8
- 230000033115 angiogenesis Effects 0.000 claims abstract 10
- 229920002678 cellulose Polymers 0.000 claims abstract 10
- 239000001913 cellulose Substances 0.000 claims abstract 10
- 235000018102 proteins Nutrition 0.000 claims description 98
- 239000000047 product Substances 0.000 claims description 95
- 210000000988 bone and bone Anatomy 0.000 claims description 78
- 239000003795 chemical substances by application Substances 0.000 claims description 19
- 235000005911 diet Nutrition 0.000 claims description 18
- 230000037213 diet Effects 0.000 claims description 18
- 239000003814 drug Substances 0.000 claims description 15
- 210000000481 breast Anatomy 0.000 claims description 13
- 230000001629 suppression Effects 0.000 claims description 10
- 102000002322 Egg Proteins Human genes 0.000 claims description 9
- 108010000912 Egg Proteins Proteins 0.000 claims description 9
- 238000000034 method Methods 0.000 claims description 9
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 claims description 8
- 235000014103 egg white Nutrition 0.000 claims description 8
- 210000000969 egg white Anatomy 0.000 claims description 8
- 230000007062 hydrolysis Effects 0.000 claims description 8
- 238000006460 hydrolysis reaction Methods 0.000 claims description 8
- 238000004519 manufacturing process Methods 0.000 claims description 8
- 238000002156 mixing Methods 0.000 claims description 8
- 230000002255 enzymatic effect Effects 0.000 claims description 6
- 108010005843 Cysteine Proteases Proteins 0.000 claims description 5
- 102000005927 Cysteine Proteases Human genes 0.000 claims description 5
- 108010022999 Serine Proteases Proteins 0.000 claims description 5
- 102000012479 Serine Proteases Human genes 0.000 claims description 5
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 claims description 4
- 235000018417 cysteine Nutrition 0.000 claims description 4
- 235000013601 eggs Nutrition 0.000 claims description 4
- 239000003531 protein hydrolysate Substances 0.000 claims description 2
- 238000000354 decomposition reaction Methods 0.000 claims 1
- 238000000605 extraction Methods 0.000 claims 1
- 230000000052 comparative effect Effects 0.000 description 31
- 239000000243 solution Substances 0.000 description 28
- 239000000203 mixture Substances 0.000 description 25
- 230000000694 effects Effects 0.000 description 22
- 210000002997 osteoclast Anatomy 0.000 description 18
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 16
- 210000004027 cell Anatomy 0.000 description 15
- 238000010612 desalination reaction Methods 0.000 description 14
- 102000011632 Caseins Human genes 0.000 description 13
- 108010076119 Caseins Proteins 0.000 description 13
- 208000020084 Bone disease Diseases 0.000 description 11
- 238000002360 preparation method Methods 0.000 description 11
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 10
- 208000001132 Osteoporosis Diseases 0.000 description 10
- 239000011575 calcium Substances 0.000 description 10
- 229910052791 calcium Inorganic materials 0.000 description 10
- 235000001465 calcium Nutrition 0.000 description 10
- 239000008367 deionised water Substances 0.000 description 10
- 229910021641 deionized water Inorganic materials 0.000 description 10
- 239000004365 Protease Substances 0.000 description 8
- 230000037182 bone density Effects 0.000 description 8
- 239000012528 membrane Substances 0.000 description 8
- 230000035755 proliferation Effects 0.000 description 8
- 239000011780 sodium chloride Substances 0.000 description 8
- 102000009840 Angiopoietins Human genes 0.000 description 7
- 108010009906 Angiopoietins Proteins 0.000 description 7
- 239000001963 growth medium Substances 0.000 description 7
- 238000001223 reverse osmosis Methods 0.000 description 7
- 235000020183 skimmed milk Nutrition 0.000 description 7
- 102000035195 Peptidases Human genes 0.000 description 6
- 108091005804 Peptidases Proteins 0.000 description 6
- 241000700159 Rattus Species 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 108010046377 Whey Proteins Proteins 0.000 description 6
- 102000007544 Whey Proteins Human genes 0.000 description 6
- 235000021240 caseins Nutrition 0.000 description 6
- 230000008859 change Effects 0.000 description 6
- 230000004069 differentiation Effects 0.000 description 6
- 235000013305 food Nutrition 0.000 description 6
- 235000019419 proteases Nutrition 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 239000003643 water by type Substances 0.000 description 6
- 208000008035 Back Pain Diseases 0.000 description 5
- 208000008930 Low Back Pain Diseases 0.000 description 5
- 229940021722 caseins Drugs 0.000 description 5
- 238000010828 elution Methods 0.000 description 5
- 230000001582 osteoblastic effect Effects 0.000 description 5
- 239000011347 resin Substances 0.000 description 5
- 229920005989 resin Polymers 0.000 description 5
- 239000013589 supplement Substances 0.000 description 5
- 235000021246 κ-casein Nutrition 0.000 description 5
- 208000010392 Bone Fractures Diseases 0.000 description 4
- 206010017076 Fracture Diseases 0.000 description 4
- 206010072132 Fracture pain Diseases 0.000 description 4
- 238000002835 absorbance Methods 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 239000008280 blood Substances 0.000 description 4
- 210000004204 blood vessel Anatomy 0.000 description 4
- 230000037118 bone strength Effects 0.000 description 4
- 125000002091 cationic group Chemical group 0.000 description 4
- 239000000796 flavoring agent Substances 0.000 description 4
- 238000004108 freeze drying Methods 0.000 description 4
- 239000000499 gel Substances 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- 229910052500 inorganic mineral Inorganic materials 0.000 description 4
- 239000003456 ion exchange resin Substances 0.000 description 4
- 229920003303 ion-exchange polymer Polymers 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 235000010755 mineral Nutrition 0.000 description 4
- 239000011707 mineral Substances 0.000 description 4
- 238000000575 proteomic method Methods 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
- 238000004659 sterilization and disinfection Methods 0.000 description 4
- 238000011282 treatment Methods 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 239000005862 Whey Substances 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 230000037396 body weight Effects 0.000 description 3
- 239000001506 calcium phosphate Substances 0.000 description 3
- 229910000389 calcium phosphate Inorganic materials 0.000 description 3
- 235000011010 calcium phosphates Nutrition 0.000 description 3
- 159000000007 calcium salts Chemical class 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 239000012141 concentrate Substances 0.000 description 3
- 235000008504 concentrate Nutrition 0.000 description 3
- 238000009826 distribution Methods 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 210000000963 osteoblast Anatomy 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 238000001556 precipitation Methods 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 3
- 239000011782 vitamin Substances 0.000 description 3
- 229930003231 vitamin Natural products 0.000 description 3
- 235000013343 vitamin Nutrition 0.000 description 3
- 229940088594 vitamin Drugs 0.000 description 3
- 235000021119 whey protein Nutrition 0.000 description 3
- 241000283690 Bos taurus Species 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 2
- 229920001661 Chitosan Polymers 0.000 description 2
- 208000034657 Convalescence Diseases 0.000 description 2
- 235000010469 Glycine max Nutrition 0.000 description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 2
- 108010046938 Macrophage Colony-Stimulating Factor Proteins 0.000 description 2
- 102100028123 Macrophage colony-stimulating factor 1 Human genes 0.000 description 2
- 108010019160 Pancreatin Proteins 0.000 description 2
- 108090000526 Papain Proteins 0.000 description 2
- 102000014128 RANK Ligand Human genes 0.000 description 2
- 108010025832 RANK Ligand Proteins 0.000 description 2
- 229920002684 Sepharose Polymers 0.000 description 2
- DRTQHJPVMGBUCF-XVFCMESISA-N Uridine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-XVFCMESISA-N 0.000 description 2
- 235000010724 Wisteria floribunda Nutrition 0.000 description 2
- 206010003246 arthritis Diseases 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 210000002798 bone marrow cell Anatomy 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 230000003185 calcium uptake Effects 0.000 description 2
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 2
- 150000001768 cations Chemical class 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 239000007979 citrate buffer Substances 0.000 description 2
- 235000020247 cow milk Nutrition 0.000 description 2
- 235000021185 dessert Nutrition 0.000 description 2
- 235000015872 dietary supplement Nutrition 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 230000007071 enzymatic hydrolysis Effects 0.000 description 2
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 2
- 230000037406 food intake Effects 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 229940088597 hormone Drugs 0.000 description 2
- 239000005556 hormone Substances 0.000 description 2
- 230000009245 menopause Effects 0.000 description 2
- 230000002503 metabolic effect Effects 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 229940055695 pancreatin Drugs 0.000 description 2
- 229940055729 papain Drugs 0.000 description 2
- 235000019834 papain Nutrition 0.000 description 2
- 230000001376 precipitating effect Effects 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 108010048734 sclerotin Proteins 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 238000006277 sulfonation reaction Methods 0.000 description 2
- 230000007306 turnover Effects 0.000 description 2
- 210000000689 upper leg Anatomy 0.000 description 2
- 150000003722 vitamin derivatives Chemical class 0.000 description 2
- 235000021247 β-casein Nutrition 0.000 description 2
- 229920000936 Agarose Polymers 0.000 description 1
- WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical compound [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 description 1
- 102000055006 Calcitonin Human genes 0.000 description 1
- 108060001064 Calcitonin Proteins 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 102000005600 Cathepsins Human genes 0.000 description 1
- 108010084457 Cathepsins Proteins 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 108010051815 Glutamyl endopeptidase Proteins 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 206010019233 Headaches Diseases 0.000 description 1
- 108060005987 Kallikrein Proteins 0.000 description 1
- 102000001399 Kallikrein Human genes 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 102000014171 Milk Proteins Human genes 0.000 description 1
- 108010011756 Milk Proteins Proteins 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 235000019482 Palm oil Nutrition 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 235000004347 Perilla Nutrition 0.000 description 1
- 244000124853 Perilla frutescens Species 0.000 description 1
- 102000003992 Peroxidases Human genes 0.000 description 1
- 108010001441 Phosphopeptides Proteins 0.000 description 1
- 101710093543 Probable non-specific lipid-transfer protein Proteins 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 108090001109 Thermolysin Proteins 0.000 description 1
- 208000009205 Tinnitus Diseases 0.000 description 1
- 229930003316 Vitamin D Natural products 0.000 description 1
- QYSXJUFSXHHAJI-XFEUOLMDSA-N Vitamin D3 Natural products C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C/C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-XFEUOLMDSA-N 0.000 description 1
- 229930003448 Vitamin K Natural products 0.000 description 1
- 238000011481 absorbance measurement Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000005030 aluminium foil Substances 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000005875 antibody response Effects 0.000 description 1
- 230000036528 appetite Effects 0.000 description 1
- 235000019789 appetite Nutrition 0.000 description 1
- 230000002917 arthritic effect Effects 0.000 description 1
- DRTQHJPVMGBUCF-PSQAKQOGSA-N beta-L-uridine Natural products O[C@H]1[C@@H](O)[C@H](CO)O[C@@H]1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-PSQAKQOGSA-N 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Substances BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 description 1
- 229910052794 bromium Inorganic materials 0.000 description 1
- 230000002308 calcification Effects 0.000 description 1
- 229960004015 calcitonin Drugs 0.000 description 1
- BBBFJLBPOGFECG-VJVYQDLKSA-N calcitonin Chemical compound N([C@H](C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N1[C@@H](CCC1)C(N)=O)C(C)C)C(=O)[C@@H]1CSSC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1 BBBFJLBPOGFECG-VJVYQDLKSA-N 0.000 description 1
- 229960005069 calcium Drugs 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- MKJXYGKVIBWPFZ-UHFFFAOYSA-L calcium lactate Chemical class [Ca+2].CC(O)C([O-])=O.CC(O)C([O-])=O MKJXYGKVIBWPFZ-UHFFFAOYSA-L 0.000 description 1
- 235000011086 calcium lactate Nutrition 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 238000002737 cell proliferation kit Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000005660 chlorination reaction Methods 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 239000002285 corn oil Substances 0.000 description 1
- 235000005687 corn oil Nutrition 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 229940099112 cornstarch Drugs 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- XQRLCLUYWUNEEH-UHFFFAOYSA-L diphosphonate(2-) Chemical compound [O-]P(=O)OP([O-])=O XQRLCLUYWUNEEH-UHFFFAOYSA-L 0.000 description 1
- 229940042399 direct acting antivirals protease inhibitors Drugs 0.000 description 1
- 238000002086 displacement chromatography Methods 0.000 description 1
- 210000003278 egg shell Anatomy 0.000 description 1
- 238000000909 electrodialysis Methods 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000002532 enzyme inhibitor Substances 0.000 description 1
- 229940125532 enzyme inhibitor Drugs 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 229940011871 estrogen Drugs 0.000 description 1
- 239000000262 estrogen Substances 0.000 description 1
- XTLNYNMNUCLWEZ-UHFFFAOYSA-N ethanol;propan-2-one Chemical compound CCO.CC(C)=O XTLNYNMNUCLWEZ-UHFFFAOYSA-N 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 235000021323 fish oil Nutrition 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 235000013312 flour Nutrition 0.000 description 1
- 235000013355 food flavoring agent Nutrition 0.000 description 1
- 238000005227 gel permeation chromatography Methods 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 231100000869 headache Toxicity 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- CJWQYWQDLBZGPD-UHFFFAOYSA-N isoflavone Natural products C1=C(OC)C(OC)=CC(OC)=C1C1=COC2=C(C=CC(C)(C)O3)C3=C(OC)C=C2C1=O CJWQYWQDLBZGPD-UHFFFAOYSA-N 0.000 description 1
- 150000002515 isoflavone derivatives Chemical class 0.000 description 1
- 235000008696 isoflavones Nutrition 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000000845 maltitol Substances 0.000 description 1
- 235000010449 maltitol Nutrition 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 230000011987 methylation Effects 0.000 description 1
- 238000007069 methylation reaction Methods 0.000 description 1
- 238000001471 micro-filtration Methods 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 235000021239 milk protein Nutrition 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 230000020395 negative regulation of osteoclast differentiation Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 235000019198 oils Nutrition 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 238000009806 oophorectomy Methods 0.000 description 1
- 230000001599 osteoclastic effect Effects 0.000 description 1
- 125000000636 p-nitrophenyl group Chemical group [H]C1=C([H])C(=C([H])C([H])=C1*)[N+]([O-])=O 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 239000002540 palm oil Substances 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 229920001277 pectin Polymers 0.000 description 1
- 239000001814 pectin Substances 0.000 description 1
- 235000010987 pectin Nutrition 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- SHUZOJHMOBOZST-UHFFFAOYSA-N phylloquinone Natural products CC(C)CCCCC(C)CCC(C)CCCC(=CCC1=C(C)C(=O)c2ccccc2C1=O)C SHUZOJHMOBOZST-UHFFFAOYSA-N 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000010926 purge Methods 0.000 description 1
- 238000007634 remodeling Methods 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- HELHAJAZNSDZJO-OLXYHTOASA-L sodium L-tartrate Chemical compound [Na+].[Na+].[O-]C(=O)[C@H](O)[C@@H](O)C([O-])=O HELHAJAZNSDZJO-OLXYHTOASA-L 0.000 description 1
- 239000001433 sodium tartrate Substances 0.000 description 1
- 229960002167 sodium tartrate Drugs 0.000 description 1
- 235000011004 sodium tartrates Nutrition 0.000 description 1
- 210000004872 soft tissue Anatomy 0.000 description 1
- 239000003549 soybean oil Substances 0.000 description 1
- 235000012424 soybean oil Nutrition 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 210000002303 tibia Anatomy 0.000 description 1
- 231100000886 tinnitus Toxicity 0.000 description 1
- DRTQHJPVMGBUCF-UHFFFAOYSA-N uracil arabinoside Natural products OC1C(O)C(CO)OC1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-UHFFFAOYSA-N 0.000 description 1
- 229940045145 uridine Drugs 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
- 235000019166 vitamin D Nutrition 0.000 description 1
- 239000011710 vitamin D Substances 0.000 description 1
- 150000003710 vitamin D derivatives Chemical class 0.000 description 1
- 235000019168 vitamin K Nutrition 0.000 description 1
- 239000011712 vitamin K Substances 0.000 description 1
- 150000003721 vitamin K derivatives Chemical class 0.000 description 1
- 239000011728 vitamin K2 Substances 0.000 description 1
- 229940046008 vitamin d Drugs 0.000 description 1
- 229940046010 vitamin k Drugs 0.000 description 1
- 235000012773 waffles Nutrition 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 235000013618 yogurt Nutrition 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/55—Protease inhibitors
- A61K38/57—Protease inhibitors from animals; from humans
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/475—Growth factors; Growth regulators
- C07K14/515—Angiogenesic factors; Angiogenin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/18—Growth factors; Growth regulators
- A61K38/1891—Angiogenesic factors; Angiogenin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/01—Hydrolysed proteins; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/22—Hormones
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/43—Enzymes; Proenzymes; Derivatives thereof
- A61K38/46—Hydrolases (3)
- A61K38/465—Hydrolases (3) acting on ester bonds (3.1), e.g. lipases, ribonucleases
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/55—Protease inhibitors
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
- A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/81—Protease inhibitors
- C07K14/8107—Endopeptidase (E.C. 3.4.21-99) inhibitors
- C07K14/8139—Cysteine protease (E.C. 3.4.22) inhibitors, e.g. cystatin
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y301/00—Hydrolases acting on ester bonds (3.1)
- C12Y301/27—Endoribonucleases producing 3'-phosphomonoesters (3.1.27)
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Immunology (AREA)
- Epidemiology (AREA)
- Gastroenterology & Hepatology (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Molecular Biology (AREA)
- Nutrition Science (AREA)
- Mycology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Physical Education & Sports Medicine (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Endocrinology (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Rheumatology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biophysics (AREA)
- Vascular Medicine (AREA)
- Animal Husbandry (AREA)
- Toxicology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The present invention relates to a kind of new protein material.The protein material include 2~15mg/100mg amount angiogenin and/or angiogenesis cellulose hydrolysate, and with the mass ratio of angiogenin and/or angiogenesis cellulose hydrolysate be 0.003~0.6 cystatin and/or cystatin hydrolysate.
Description
The application be the applying date on 07 31st, 2012, the Application No. 201280074973.9 into China, invention
The divisional application of the application of entitled " new protein material ".
Technical field
The present invention relates to new protein material, and medicine, diet product or feed comprising the protein material, and for preventing and
Treat bone disease.The protein material, which has, promotes osteoblastic proliferation, and the bone for suppressing osteoclast differentiation and osteoclast is inhaled
The function of receipts.Therefore, the protein material is used to prevent and treat various bone diseases, such as osteoporosis, fracture, rheumatism and pass
Section is scorching.
Background technology
In recent years, in the world along with social senilization etc., various bone diseases, such as osteoporosis, fracture and
Low Back Pain increase, and have turned into serious social concern.These diseases are declined by calcium insufficiency of intake, calcium uptake ability, menopause
Hormone imbalances etc. cause afterwards.It is thought that begin through activation Gegenbaur's cell and bon e formation from life early stage increases body as much as possible
Interior bone amount, and increase maximum bone amount and bone strength (bone density+sclerotin) and preventing various bone diseases, such as osteoporosis, bone
It is effective in terms of folding and Low Back Pain.Notice that term " sclerotin " is related to bone microstructure, metabolic turnover (metabolic
Turnover), micro-crack and calcification.It is said that various bone diseases, such as osteoporosis, fracture and Low Back Pain can be by suppressing osteoclastic thin
The bone information of born of the same parents prevents.Bone absorbs in a balanced fashion and forms (remodeling) repeatedly.However, hormone is put down after due to menopause
When weighing apparatus change etc. causes the bone information to exceed bon e formation, various bone diseases, such as osteoporosis, fracture and Low Back Pain can occur.Therefore,
By suppressing the bone information of osteoclast and maintaining bone strength bone can be made to be strengthened in constant level.
In view of said circumstances, in order to strengthen bone, individually addition calcium salt, such as calcium carbonate, calcium phosphate or breast intake of
The calcium salt of sour calcium or natural calcium product, such as whey calcium, the medicine of bovine bone powder or egg shell, diet product or feed etc..Also make
With comprising such calcium product and with calcium uptake facilitation effect material, such as the medicine of casein phosphopeptide or oligosaccharide,
Diet product or feed etc. strengthen bone.However, calcium absorptivity is when the diet product that intake includes calcium salt or natural calcium product
Less than 50%, and the most of calcium absorbed may be excreted without being absorbed.In addition, even if calcium is absorbed into vivo,
Because the compatibility to bone can be different according to the type of its form or the nutritional ingredient absorbed simultaneously, Bone m etabolism may not be also shown
Improvement or bone strengthen effect.As for treating osteoporosis or strengthening estrogen preparations known to the medicine of bone, activity dimension
Raw plain D3Product, vitamin K2Product, diphosphonate product and calcitonin product etc., and it is for example anti-to also developed newtype drug
RANKL antibody.However, these medicines can have side effect such as tinnitus, headache or poor appetite.In addition, from safety and cost
Etc. viewpoint, above-mentioned substance is in the situation that can not be added to diet product at present.Therefore, exploitation has been needed according to various bone diseases
The case such as property of osteoporosis, fracture and Low Back Pain, can long-term oral uptake, by promoting bone growing and suppress bone information and increase
Add bone strength, and prevention can be desired to have or treat such bone hardening agent, diet product or the feed of the effect of various bone diseases.
Several food materials for being intended to improve bone strength be present, for example, having reported the alkaline protein or its enzymatic from breast
Peptide fraction display activity of osteoblast proliferation, the bone information inhibitory activity of osteoclast of hydrolysate, and bone thus are strong
Change is acted on (referring to patent document 1).Also have reported the angiogenin included in the basic protein fraction from breast and suppression half
Cysteine proteases albumen independently has the function of improving Bone m etabolism (referring to patent document 2 and 3).
Prior art literature
Patent document
[patent document 1] JP-A-H08-151331
[patent document 2] JP-A-H10-7585
[patent document 3] JP-A-2000-281587
The content of the invention
Problems to be solved by the invention
The purpose of the present invention promotes skeletonization to provide new protein material, the protein material safety by daily ingestion
Cell is bred, while suppresses the bone information of osteoclast differentiation and osteoclast, thus can strengthen bone.
Another object of the present invention is used to prevent and treat various bone diseases such as osteoporosis to provide by oral uptake
Disease, fracture, rheumatism and arthritic bone strengthen medicine, diet product or feed.
The solution used to solve the problem
It has been found by the present inventors that effectively facilitate osteoblastic proliferation and suppress the bone of osteoclast differentiation and osteoclast
The effect of absorption can include the hydrolysate of specified quantitative angiogenin and/or angiogenin by absorbing, and relative to blood
The hydrolysate of pipe generation element and/or angiogenin further includes cystatin with specific mass ratio
(cystatin) and/or the protein material of the hydrolysate of cystatin obtains.The discovery causes this hair
Bright completion.
Specifically, the present invention includes following aspect：
(1) a kind of protein material, it includes the angiogenin of 2~15mg/100mg amount and/or angiogenin
Hydrolysate, and using with the mass ratio of angiogenin and/or the hydrolysate of angiogenin as 0.003~0.6 suppression
The hydrolysate of cysteine protease protein and/or cystatin.
(2) a kind of diet product or feed, it includes the protein material according to (1).
(3) a kind of bone hardening agent, it is comprising the protein material described in basis (1) as active component.
(4) a kind of bone intensifying method, it includes the protein material according to (1) with the amount intake of more than 5mg/ days.
(5) method that one kind prepares the protein material according to (1), 1)~3 it comprises the following steps)：
1) hydrolysate of angiogenin and/or angiogenin is prepared；
2) hydrolysate of cystatin and/or cystatin is prepared；With
3) mixing according to it is above-mentioned 2) described in cystatin and/or cystatin
Hydrolysate and according to it is above-mentioned 1) described in angiogenin and/or angiogenin hydrolysate, with cause press down half Guang
The hydrolysate of serine protease albumen and/or cystatin and angiogenin and/or angiogenin
Hydrolysate mass ratio be 0.003~0.6.
(6) method of protein material of a kind of manufacture according to (1), it is included from breast and/or the material from breast carries
Take hydrolysate and cystatin comprising angiogenin and/or angiogenin and/or half Guang of suppression
The fraction of the hydrolysate of serine protease albumen, to cause cystatin and/or suppression cysteine protein
The hydrolysate of zymoprotein is 0.003~0.6 with the mass ratio of angiogenin and/or the hydrolysate of angiogenin.
(7) according to the method described in (6), its further comprise the angiogenin being included in the fraction and/or
The other step of cystatin enzymatic hydrolysis.
The effect of invention
The protein material of the present invention is by promoting osteoblastic proliferation and suppressing osteoclast differentiation and osteoclast
Bone information and show significant bone invigoration effect.Medicine, diet product or the feed enhancement bone of the present invention, and for preventing and treating
Various bone diseases, such as osteoporosis, fracture, rheumatism and arthritis.
Embodiment
The protein material of the present invention is characterised by that the protein material includes the angiogenin and/or blood vessel of specified quantitative
The hydrolysate of element is generated, and is entered relative to the hydrolysate of angiogenin and/or angiogenin with extra fine quality ratio
One step includes the hydrolysate of cystatin and/or cystatin.
The protein material of the present invention can be by including angiogenin and/or angiogenin than mixing with extra fine quality
Hydrolysate fraction and comprising cystatin and/or cystatin hydrolysis production
The mixture that the fraction of thing obtains, or by being extracted from breast or from newborn material such as skimmed milk or whey etc. with extra fine quality
Than the hydrolysate comprising angiogenin and/or angiogenin and cystatin and/or half Guang of suppression
The fraction of the hydrolysate of serine protease albumen is come material for preparing etc..The protein material of the present invention, which may also include, passes through enzymatic
Hydrolyze material prepared by angiogenin and/or cystatin.
When by mixing the fraction of the hydrolysate comprising angiogenin and/or angiogenin and comprising pressing down half Guang
The fraction of the hydrolysate of serine protease albumen and/or cystatin come prepare the present invention protein material
When, by the fraction of the breast preparation of mammal such as people, milk cow, buffalo, goat or sheep, the fraction produced by genetic engineering,
Or graded by the level of blood or internal organs purifying and can be used as the hydrolysate comprising angiogenin and/or angiogenin
The fraction of fraction and hydrolysate comprising cystatin and/or cystatin.
The angiogenin or cystatin reagent for the purifying being obtained commercially can be used.In this case, it is of the invention
Protein material can by adjust the hydrolysate of cystatin and/or cystatin with
It is prepared by the mass ratio of the hydrolysate of angiogenin and/or angiogenin.
Tried using more than one protease by the above-mentioned fraction comprising angiogenin of enzymatic hydrolysis, angiogenin
The product of the acquisitions such as agent, the fraction comprising cystatin or cystatin reagent can be used as
The hydrolysate of angiogenin or the hydrolysate of cystatin.
When by from breast or from breast material such as skimmed milk or whey directly extract include extra fine quality than blood vessel life
Into element and/or the hydrolysate and cystatin and/or suppression cysteine proteinase egg of angiogenin
When the fraction of white hydrolysate is to prepare the protein material of the present invention, for example, breast or the material from breast can be handed over cation
Change resin to be in contact, the milk protein being then adsorbed on resin can elute under 0.1~2.0M salinity, use counter-infiltration
Desalination and the concentrations such as film, electrodialysis film, milipore filter or microfiltration membranes, protease such as trypsase, pancreatin, pancreas are optionally used afterwards
Chrymotrypsin, pepsin, papain, kallikrein, cathepsin, thermolysin or V8 protease enter
The limited molecular weight for being hydrolyzed into less than 8000 of row.When carrying out limited hydrolysis using protease, the lower limit of preferred molecular weight is
More than 500.Thus obtained protein material can be dried by being freeze-dried or being spray-dried etc..
When making the protein material of the present invention carry out LC/MS/MS analyses, digestive ferment is being used in the usual way in reduction bar
Modified under part and limited hydrolysis, so as to carry out the Proteomic analysis of protein material, afterwards, it was demonstrated that protein material includes extremely
A kind of few protein, such as α s1- caseins, α s2- caseins, beta-casein or κ-casein, and except angiogenin
And/or the hydrolysate and cystatin and/or cystatin of angiogenin
Its protein hydrolysate beyond hydrolysate.
The protein material of the present invention includes the 2~15mg/100mg angiogenin of amount and/or the water of angiogenin
Product is solved, and is included relative to the hydrolysate of angiogenin and/or angiogenin with 0.003~0.6 mass ratio
The hydrolysate of cystatin and/or cystatin.
As shown in following test examples, when cystatin and/or the water of cystatin
When the mass ratio for solving the hydrolysate of product and angiogenin and/or angiogenin is 0.003~0.6, bone invigoration effect
Than individually intake angiogenin and/or angiogenin hydrolysate or cystatin and/or
The situation of the hydrolysate of cystatin more effectively obtains.
Pay attention to, it is only for reference, it is about in the content of cow's milk angiopoietin and/or the hydrolysate of angiogenin
0.001%, and the hydrolysate and blood of cystatin and/or cystatin in cow's milk
The mass ratio of the hydrolysate of pipe generation element and/or angiogenin is about 2.Given birth in whey protein concentrate (WPC) medium vessels
Content into element and/or the hydrolysate of angiogenin is about 0.1%, and presses down cysteine egg in whey protein concentrate
The hydrolysis of the hydrolysate and angiogenin and/or angiogenin of white zymoprotein and/or cystatin
The mass ratio of product is about 3.
The protein material of the present invention can be prepared as bone hardening agent by being properly added protein material as active component.This
The protein material of invention can be directly used as bone hardening agent.When being formulated as bone hardening agent, it can mix and be generally used for medicine, diet product
With the raw material of feed etc., such as carbohydrate, lipid, proteins,vitamins,minerals or flavouring agent, and can be with usual way
It is configured to powdery medicine, granule, tablet, capsule or drinkable preparation etc..The protein material of the present invention can be with also showing
Show that other compositions such as calcium, vitamin D, vitamin K or the isoflavones of bone invigoration effect are used together.
As shown in following zooperies, the protein material of the present invention can when with the amount oral uptake of more than 5mg/kg body weight
Strengthen bone.Because the intake of the experimental animal corresponds to the intake of adult in terms of blood concentration (referring to Mitsuyoshi
Nakajima (1993), " Yakkou Hyoka Vol.8 ", Hirokawa-Shoten Ltd., pp.2-18), it is expected to obtain bone
Invigoration effect, it is various to prevent or treat particularly through adult more than the daily ingestion 5mg of one people one protein materials of the invention
Bone disease such as osteoporosis, fracture, rheumatism and arthritis.Therefore, when being mixed with bone hardening agent etc., protein material can add
Enter wherein to take in above-mentioned necessary amount.
The protein material of the present invention can add full diet product, such as Yoghourt, beverage, Waffle (wafer) or dessert
(dessert) in.In this case, protein material of the invention preferably according to the form of diet product with 0.25~1000mg/
The amount addition of 100g diet products.It is expected that bone invigoration effect can be by keeping above-mentioned combined amount to obtain.The protein material of the present invention
Feed can be also added, such as animal feeding-stuff or pet food, so as to prepare bone fortified feed.In such a situation it is preferred that with 0.25~
The protein material of the amount addition present invention of 1000mg/100g feeds.
When being prepared in the form of medicine, diet product or feed and using protein material of the invention, albumen of the invention
Material can be used by being suspended or dissolved in deionized water and being stirred.Stirring/mixing condition is not particularly limited, as long as
Protein material uniformly mixes.The stirrings such as ultra-dispersed device or TK mixer for well-distribution also can be used to mix.
The solution of protein material can optionally employ the desalinations such as reverse osmosis membrane or concentration, or freeze-drying so that solution can hold
It is easy for medicine, diet product or feed.
The protein material of the present invention is demonstrate especially that, is carried out even in protein material in medicine, diet product or Feed Manufacturing
In commonly use sterilization treatment when also maintain bone strengthen activity.When protein material is used as powder, protein material, which can be dried, to be added
Heat sterilization.The present invention protein material can in a variety of manners as liquid, gel, powder or particle be used for medicine, diet product or
Feed.
Below the present invention is further described more fully by reference example, embodiment and test example.Pay attention to, following implementation
Example is merely intended to illustration purpose, and should not be construed as limiting the present invention.
Reference example 1
The preparation (1) of angiogenin fraction
Post (sulfonation Chitopearl filled with 30kg cationic ion-exchange resins；By Fuji Spinning Co., Ltd.
Manufacture) fully washed with deionized water, then 1000 liters of unpasteurized skimmed milks (pH6.7) are placed in post.Use deionized water
After abundant column scrubber, 0.1~2.0M sodium chloride elution of the protein linear gradient of absorption.Elution comprising angiogenin
Fraction is classified using S- Sepharose cations displacement chromatography (being manufactured by Amersham Bioscientific), and gained includes blood vessel
The fraction of generation element heats 10 minutes at 90 DEG C, and is centrifuged off precipitating.Fraction comprising angiogenin is further
Continue gel permeation chromatography (post：Superose 12).Gained eluate uses reverse osmosis membrane desalination, and by the eluate of desalination
Freeze-drying, so as to obtain 16.5g angiogenin fraction, its angiogenin purity is 90%.These continuous operation weights
It is multiple 30 times.
Reference example 2
The preparation (2) of angiogenin fraction
Post (being manufactured by GE Healthcare) filled with 10kg heparin-agaroses is fully washed with deionized water, then
500 liters of unpasteurized skimmed milks (pH6.7) are placed in post.After 0.5M sodium chloride solution column scrubbers, the protein of absorption is used
1.5M sodium chloride solutions elute.Eluate uses reverse osmosis membrane desalination, and the eluate of desalination is freeze-dried, so as to obtain
18g angiogenin fraction, its angiogenin purity are 5%.Above-mentioned continuous operation repeats 50 times.
Reference example 3
The preparation of cystatin fraction
100,000 liter 5% of lactoalbumin soln heats 10 minutes at 90 DEG C, and is centrifuged off precipitating.Post is filled out
Filled with by the way that carboxy methylation papain and Tresyl-Toyopearl (being manufactured by Tosoh Corporation) are combined into system
Standby carrier.After being balanced with 0.5M sodium chloride solutions, above-mentioned lactoalbumin soln is placed in post.Then, by post 0.5M chlorinations
Sodium solution and the 0.5M sodium chloride solution sequential purges containing polysorbas20 (0.1%).Afterwards, comprising cystatin
Fraction with 20mM acetic acid -0.5M sodium chloride solutions elute.The fraction of elution is neutralized with 1M sodium hydroxide solutions immediately.Eluate
Then reverse osmosis membrane desalination, the eluate freeze-drying of desalination, so as to obtain 9.6g cystatin are used
Fraction, there is cystatin purity 90%.Above-mentioned continuous operation is repeated 20 times.
Embodiment 1
Mix and obtained in the angiogenin obtained in 5 points 30 milligrams of (5.30mg) reference examples 1,84.67mg reference examples 2
The angiogenin fraction obtained, and the cystatin fraction obtained in 0.03mg reference examples 3, so as to prepare egg
White material (embodiment product 1), the content of the hydrolysate of its angiopoietin and/or angiogenin is 10mg/
The hydrolysate of 100mg, cystatin and/or cystatin and angiogenin and/
Or the mass ratio of the hydrolysate of angiogenin is 0.003.
Embodiment 2
Mix and obtained in the angiogenin obtained in 5 points 35 milligrams of (5.35mg) reference examples 1,83.65mg reference examples 2
The angiogenin fraction obtained, and the cystatin fraction obtained in 1.00mg reference examples 3, so as to prepare egg
White material (embodiment product 2), the content of the hydrolysate of its angiopoietin and/or angiogenin is 10mg/
The hydrolysate of 100mg, cystatin and/or cystatin and angiogenin and/
Or the mass ratio of the hydrolysate of angiogenin is 0.1.
Embodiment 3
Mix and obtained in the angiogenin obtained in 5 points 65 milligrams of (5.65mg) reference examples 1,78.35mg reference examples 2
The angiogenin fraction obtained, and the cystatin fraction obtained in 6.00mg reference examples 3, so as to prepare egg
White material (embodiment product 3), the content of the hydrolysate of its angiopoietin and/or angiogenin is 10mg/
The hydrolysate of 100mg, cystatin and/or cystatin and angiogenin and/
Or the mass ratio of the hydrolysate of angiogenin is 0.6.
[comparative example 1]
Mix and obtained in the angiogenin obtained in 5 points 30 milligrams of (5.30mg) reference examples 1,84.68mg reference examples 2
The angiogenin fraction obtained, and the cystatin fraction obtained in 0.02mg reference examples 3, so as to prepare egg
White material (comparative example product 1), the content of the hydrolysate of its angiopoietin and/or angiogenin is 10mg/
The hydrolysate of 100mg, cystatin and/or cystatin and angiogenin and/
Or the mass ratio of the hydrolysate of angiogenin is 0.002.
[comparative example 2]
Mix and obtained in the angiogenin obtained in 5 points 68 milligrams of (5.68mg) reference examples 1,77.82mg reference examples 2
The angiogenin fraction obtained, and the cystatin fraction obtained in 6.50mg reference examples 3, so as to prepare egg
White material (comparative example product 2), the content of the hydrolysate of its angiopoietin and/or angiogenin is 10mg/
The hydrolysate of 100mg, cystatin and/or cystatin and angiogenin and/
Or the mass ratio of the hydrolysate of angiogenin is 0.65.
[test example 1]
Determine osteoblastic proliferation effect, the bone information of osteoclast of embodiment product 1~3 and comparative example product 1 and 2
Inhibition and osteoclast differentiation inhibition.
Osteoblastic proliferation effect determines as described below.The cell line (MC3T3-E1) of Gegenbaur's cell is with 2 × 103Cell/
The concentration in hole is inoculated in 96 porocyte culture plates, and using supplement have 10% hyclone (FBS) α-MEM culture mediums (by
GIBCO is manufactured) cultivate 24 hours.After culture medium removes completely, by the 90 μ l α-MEM culture mediums without FBS, and 10 μ l are included
Embodiment product 1~3 and 1 and 2 any solution of comparative example product are added to each hole.Cell is further cultivated 24 hours.
Addition is contained in cell proliferation reagent box (Cell Proliferation Kit, manufactured by GE Healthcare) bromine deoxidation
After uridine (BrdU), by cell culture 2 hours, and with the anti-BrdU antibody responses of peroxidase labelling.3 are being added,
3 ', 5, after 5 '-tetramethyl benzidine (substrate), activity of osteoblast proliferation passes through by the absorbance measurement under measure 450nm
The amount that BrdU introduces cell determines.When the absorbance under 450nm is significantly higher than wherein embodiment product 1~3 and comparative example system
When product 1 and 2 are not added to the absorbance under the 450nm of the group (control) of culture medium, activity of osteoblast proliferation is defined as sun
Property.
The inhibition of the bone information of osteoclast determines as described below.Shin bone and femur are taken out from rabbit (5 days big).Remove
After removing soft tissue, by the chopping of these bones machinery and by the full bone marrow cell comprising osteoclast be scattered in supplement have 5%FBS α-
In MEM culture mediums, then with 1 × 106The concentration of cells/well is inoculated in the hole of crystallinity calcium phosphate plate (being manufactured by Corning)
On.Culture medium is removed completely, and 180 μ l supplement is had to 5%FBS α-MEM culture mediums within 2 hours after culture is started, and
20 μ l's is added to each hole comprising embodiment product 1~3 and 1 and 2 any solution of comparative example product.Cell culture 72 is small
When.After removing cell by the liquor natrii hypochloritis of addition 5%, it is formed at using stereoscope shooting on the hole of calcium phosphate plate
Absorption nest (pits), and its area is determined by graphical analysis, so that it is determined that the inhibition of the bone information of osteoclast
(Takeshi Seno etc., " Manual of selected cultured cell lines for bioscience
biotechnology”,pp.199-200,1993).When nest area is significantly less than wherein embodiment product 1~3 and comparative example system
When product 1 and 2 are not added to the area of group (control) of culture medium, sun is defined as to the inhibitory activity of the bone information of osteoclast
Property.
The inhibition of osteoclast differentiation determines as described below.From the femur collection of ddy mouse (7 or 8 weeks big, male)
Bone marrow cell with 4 × 104The concentration of cells/well is inoculated on 96 orifice plates, and in 37 DEG C and 5%CO2Under 200 μ l supplement have
Cultivated in 10%FBS and M-CSF (25ng/ml) α-MEM culture mediums.Culture medium is removed completely within 2 hours after culture is started,
And 180 μ l supplement is had to 10%FBS, RANKL (5ng/ml) and M-CSF (25ng/ml) α-MEM culture mediums, and 20 μ l
Each hole is added to comprising embodiment product 1 to 3 and 1 and 2 any solution of comparative example product, and in 37 DEG C and 5%CO2Condition
Lower culture cell 2 days.After culture medium is changed, cell is further cultivated 1 day.After the completion of culture, nutrient solution is removed, uses PBS
Washing, and with acetone-ethanol (1:1) solution handles 1 minute to fix cell.Afterwards, 1.5mg/ml p-nitrophenyl is added
Disodic alkaliine -20mM sodium tartrate -50mM citrate buffer solutions (pH 4.5) (100 μ l/ holes), and 30 points are reacted at room temperature
Clock, it is subsequently added into 1M sodium hydroxide solutions (50 μ l/ holes) terminating reaction.The absorbance under 405nm is determined, as osteoclast point
The index of change/mutation.When the absorbance under addition embodiment product 1 to 3 or the 405nm of the group of comparative example product 1 or 2 is significantly low
The extinction not being added in wherein embodiment product 1 to 3 and comparative example product 1 and 2 under the 405nm of the group (control) of culture medium
When spending, the positive is defined as to the inhibitory activity of osteoclast differentiation.
As a result it is shown in table 1.
Table 1
As shown in table 1, sun is shown in all raji cell assay Rajis corresponding to the embodiment product 1 to 3 of protein material of the present invention
Property activity.Comparative example product 1 and 2 also shows positive active in some raji cell assay Rajis, but has a raji cell assay Raji display negative
Activity.
Embodiment 4
(sulfonation Chitopearl, manufactured filled with 400g cationic ion-exchange resins by Fuji Spinning Co., Ltd.s)
Post (diameter：4cm, height：30cm) fully washed with deionized water, and by 40 liters of unpasteurized skimmed milks (pH 6.7) with
25ml/min flow is placed in post.After fully being washed with deionized water, the albumen that is adsorbed on resin is using containing 0.78M chlorine
Change 0.02M carbonate buffer solutions (pH 7.0) elution of sodium.Eluate uses reverse osmosis membrane desalination, and the eluate of desalination is cold
It is lyophilized dry, so as to obtain 18g powdered proteins material (embodiment product 4).Protein material includes blood vessel of the amount for 2mg/100mg
The hydrolysate of generation element and/or angiogenin, cystatin and/or cystatin
Hydrolysate and angiogenin and/or the mass ratio of hydrolysate of angiogenin be 0.5.Protein material can be used directly
Make the active component of bone hardening agent or bone hardening agent.As the result of Proteomic analysis, it is found that protein material includes β-junket egg
White hydrolysate and the hydrolysate of κ-casein.
Embodiment 5
Filled with 30kg cationic ion-exchange resins (SP Toyopearl；Manufactured by Tosoh Corporation) post it is (straight
Footpath：20cm, height：100cm) fully washed with deionized water, and by the heat sterilization 3t wheys (pH of 15 minutes at 75 DEG C
6.2) it is placed in 10l/min flow velocity in post.After fully being washed with deionized water, the protein use being adsorbed on resin contains
0.1M citrate buffer solutions (pH 5.7) elution of 0.68M sodium chloride.Eluate uses reverse osmosis membrane desalination, and washing desalination
De- thing freeze-drying.Above-mentioned continuous operation is repeated 20 times, so as to obtain 3.3kg powdered proteins material (embodiment product 5).Egg
White material includes amount as 15mg/100mg angiogenin and/or the hydrolysate of angiogenin, presses down cysteine protein
The hydrolysis of the hydrolysate of zymoprotein and/or cystatin and angiogenin and/or angiogenin is produced
The mass ratio of thing is 0.01.Protein material can be directly used as the active component of bone hardening agent or bone hardening agent.As protein group
The result of analysis, it is found that protein material includes the hydrolysate of α s1- caseins and κ-casein.
Embodiment 6
The protein material of four grams of (4g) embodiment products 4 is dissolved in 800ml water.Pancreatin is added with 0.02wt% final concentration
After (being manufactured by Sigma) (it is protease), mixture is then subjected to enzymatic treatment 8 hours in 37 DEG C.At 90 DEG C of heat
After reason inactivates 5 minutes zymoprotein, mixture is freeze-dried, so as to obtain 3.2g protein material (embodiment product 6).By
This protein material obtained includes hydrolysate of the amount for 2.0mg/100mg angiogenin, protease inhibitors hydrolysis production
The mass ratio of the hydrolysate of thing and angiogenin is 0.45, and the molecular weight of protein material is less than 8000.Therefore, albumen material
Material can be directly used as the active component of bone hardening agent or bone hardening agent.As the result of Proteomic analysis, protein material is found
Hydrolysate comprising beta-casein and κ-casein.
Embodiment 7
The protein material of four grams of (4g) embodiment products 5 is dissolved in 800ml water.Add trypsase (being manufactured by Sigma)
After (it is protease) is to obtain 0.03wt% final concentration, mixture is subjected to enzymatic treatment 8 hours in 37 DEG C.Pass through 90
After DEG C heat treatment inactivates 5 minutes zymoprotein, mixture is freeze-dried, so as to obtain 3.0g protein material (embodiment system
Product 7).Thus obtained protein material includes hydrolysate of the amount for 14mg/100mg angiogenin, egg in protein material
The mass ratio of the hydrolysate of white enzyme inhibitor hydrolysate and angiogenin is 0.015, and the molecular weight of protein material is
Less than 8000.Therefore, protein material can be directly used as the active component of bone hardening agent or bone hardening agent.As Proteomic analysis
Result, it is found that protein material includes the hydrolysate of α s1- caseins and κ-casein.
[comparative example 3]
The cystatin fraction and 100mg embodiment systems that will be obtained in ten milligrams of (10mg) reference examples 3
The protein material mixing of product 4, so as to prepare protein material (comparative example product 3), its angiopoietin and/or angiogenin
The content of hydrolysate be 1.8mg/100mg, cystatin and/or cystatin
Hydrolysate is 5 with the mass ratio of angiogenin and/or the hydrolysate of angiogenin.
[comparative example 4]
The protein material mixing of the angiogenin fraction and 2g embodiments product 5 that are obtained in one gram of (1g) reference example 1 is simultaneously
It is dissolved in 800ml water.Using after 0.02wt% final concentration addition trypsase (being manufactured by Sigma) (it is protease),
Mixture is subjected to enzymatic treatment 12 hours in 37 DEG C.It is after inactivating 5 minutes zymoprotein by 90 DEG C of heat treatments, mixture is cold
It is lyophilized dry, so as to obtain 2.8g protein material (comparative example product 4).It is 39mg/ that thus obtained protein material, which includes amount,
The hydrolysate of 100mg angiogenin, the hydrolysis of the hydrolysate and angiogenin of cystatin
The mass ratio of product is 0.0025.
Comparative example 5
Filled with 100g cationic ion-exchange resins (CM Sepharose FF；Manufactured by GE Healthcare) post it is (straight
Footpath：5cm, height：5cm) fully washed with deionized water, and by 40 liters of unpasteurized skimmed milks (pH 6.7) with 40ml/min's
Flow is placed in post.After fully being washed with deionized water, the protein being adsorbed on resin uses the 0.2M of the sodium chloride containing 0.98M
Carbonate buffer solution (pH6.8) elutes.Eluate uses reverse osmosis membrane desalination, and the eluate of desalination is freeze-dried.It is above-mentioned
Continuous operation is repeated 20 times, so as to obtain 20g powdered proteins material (comparative example product 5).Protein material is comprising amount
1.5mg/100mg angiogenin and/or the hydrolysate of angiogenin, cystatin and/or suppression
The mass ratio of the hydrolysate of cysteine protease protein and angiogenin and/or the hydrolysate of angiogenin is
0.001。
Test example 2
Each bone invigoration effect of embodiment product 4 and 5 and comparative example product 3 and 5 is determined by zoopery.C3H/
HeJ mouse (5 weeks big, male) are used for zoopery.After 1 week adapts to environment, mouse is divided into five groups (6 mouse/groups).Mouse
Using test tube with every 1kg body weight 5mg amount oral administration embodiment product 4 or 5 or comparative example product 3 or 5, once a day,
Totally 4 weeks.Control group does not administer any embodiment product 4 and 5, does not administer comparative example product 3 and 5 yet.The (the 4th after completing to administer
Week), use the bone density of the right chamber bone of each mouse of micro- CT (being manufactured by Rigaku Corporation) measure.As a result it is shown in table 2.
Table 2
| Bone density (mg/cm3) | |
| Control group | 1299±10 |
| Embodiment product 4 | 1328±11 |
| Embodiment product 5 | 1331±12 |
| Comparative example product 3 | 1302±10 |
| Comparative example product 5 | 1303±9 |
As shown in table 2, oral administration is the group of the embodiment product 4 or 5 of protein material of the present invention, with control group and orally
The group of administering comparative example product 3 or 5 is compared, and shows the bone density dramatically increased.
Test example 3
Each bone invigoration effect of embodiment product 6 and 7 and comparative example product 4 and 5 is determined by zoopery.48
SD rats (51 weeks big, female) are used for zoopery.
Rat is divided into six groups (8 rat/groups).Five groups of carry out oophorectomies, and by remaining one group of carry out sham-operation
(sham surgery).After 4 week convalescence, ovariectomized rat is carried out using pipe with every 1kg rat body weights 5mg amount
Oral administration embodiment product 6 or 7 or comparative example product 4 or 5, once a day, totally 16 weeks.Control group does not take embodiment system
Product 6 and 7 and comparative example product 4 and 5 it is any.After 4 week convalescence, carry out the rat of sham-operation with control group identical
Mode feeding 16 weeks.After completing to be administered (the 16th week), determined using micro--CT (being manufactured by Rigaku Corporation) major
The bone density of the right chamber bone of mouse.As a result it is shown in table 3.
Table 3
| Bone density (mg/cm3) | |
| Control group | 550±10 |
| Sham-operation group | 600±9 |
| Embodiment product 6 | 597±11 |
| Embodiment product 7 | 595±12 |
| Comparative example product 4 | 556±13 |
| Comparative example product 5 | 554±11 |
As shown in table 3, oral administration is the group of the embodiment product 6 or 7 of protein material of the present invention, with control group and orally
The group of administering comparative example product 4 or 5 is compared, and shows the bone density dramatically increased.In addition, the bone of bone density close to sham-operation group is close
Degree.
Embodiment 8
Bone strengthens the preparation of liquid nutritional supplement
The protein material of five grams of (5g) embodiment products 4 is dissolved in 4995g deionized waters.Use TK- mixer for well-distribution (TK
ROBO MICS；Manufactured by Tokushu Kika Kogyo co., ltd.) agitating solution 30 minutes under 6000rpm, so as to obtain
The solution of 100mg/100g embodiment product 4 must be included.Then, 4.0kg caseins, 5.0kg soybean are added to 5.0kg solution
Albumen, 1.0kg fish oil, 3.0kg perilla oils, 18.0kg dextrin, 6.0kg mineral mixtures, 1.95kg vitamin mixtures,
2.0kg emulsifying agents, 4.0kg stabilization agents and 0.05kg essence.Fill this blend into retort pouch (retort pouch) (200ml)
In and use Sterilization Kettle (retort sterilizer) (1 type III pressure vessel, RCS-4CRTGN；By Hisaka Works, Ltd.
Manufacture) sterilized 20 minutes at 121 DEG C, strengthen liquid nutritional supplement so as to produce 50kg bone.Any precipitation is observed, and
Do not feel off-flavor in thus obtained bone strengthens liquid nutritional composition.
Embodiment 9
Bone strengthens the preparation of gel sample food
The protein material of two grams of (2g) embodiment products 5 is dissolved in 708g deionized waters.Use ultra-dispersed device (ULTRA-
TURRAX T-25；Manufactured by IKA Japan) agitating solution 30 minutes under 9500rpm.40g D-sorbites, 2g acids, 2g
Essence, 5g pectin, 5g whey protein concentrates, 1g calcium lactates and 235g deionized waters are added in solution., will after stirring and mixing
Mixture is fitted into 200ml In Aluminium Foil Packings, and is sterilized 20 minutes at 85 DEG C, by package encapsulation, so as to obtain five bags (200g)
Bone strengthens gel sample food.Any precipitation is observed, and does not feel different in thus obtained bone strengthens gel sample food
Normal flavor.
Embodiment 10
Bone strengthens the preparation of drink
Two grams of (2g) acidulants are dissolved in 706g deionized waters, and the protein material of 4g embodiments product 6 is dissolved in into solution
In.Use ultra-dispersed device (ULTRA-TURRAX T-25；Manufactured by IKA Japan) solution 30 is stirred under 9500rpm divides
Clock.After addition 100g maltitols, 20g reduction starch syrup, 2g essence and 166g deionized waters, fill this blend into
In 100ml vials.After being sterilized 15 seconds at 95 DEG C, by bottle tight seal, strengthen drink so as to obtain ten bottles of (100ml) bones.See
Any precipitation is examined, and does not feel off-flavor in thus obtained bone strengthens drink.
Embodiment 11
The preparation of bone fortified feed
The protein material of two grams of (2kg) embodiment products 7 is dissolved in 95kg deionized waters.Use TK- mixer for well-distribution
(MARK II 160；Manufactured by PRIMIX Corporation) solution is stirred under 3600rpm 40 minutes, so as to obtain
The solution of embodiment product 7 comprising 2g/100g.Then, by 12kg soy meals, the powdered skimmed milks of 14kg, 4kg soybean oils,
2kg corn oils, 23.2kg palm oils, 14kg cornstarch, 9kg flour, 2kg rice brans, 5kg vitamin mixtures, 2.8kg fibers
Element and 2kg mineral mixtures are added in 10kg solution.It will be sterilized 4 minutes at 120 DEG C of mixture, it is strong so as to obtain 100kg bones
Change dog food.
Embodiment 12
The preparation of bone hardening agent (tablet)
Raw material is mixed with the ratio shown in table 4.Then, shape in a usual manner and ingot 1g mixtures processed, so as to prepare
Bone hardening agent.
Table 4
| Water-containing crystal glucose | 92.5% (wt%) |
| Protein material (embodiment product 1) | 1.0% |
| Mineral mixture | 5.0% |
| Sugar ester | 1.0% |
| Essence | 0.5% |
Claims (7)
1. a kind of protein material, it includes the angiogenin and/or angiogenesis cellulose hydrolysate of 2~15mg/100mg amount
With with the mass ratio of angiogenin and/or angiogenesis cellulose hydrolysate be 0.003~0.6 suppression cysteine proteinase egg
White and/or cystatin hydrolysate, the angiogenesis cellulose hydrolysate and the suppression cysteine egg
The molecular weight of white zymoprotein hydrolysate is 500-8000.
2. a kind of diet product or feed, it includes protein material according to claim 1.
3. a kind of bone hardening agent, it includes protein material according to claim 1 as active component.
4. protein material according to claim 1 is being prepared by the purposes in medicament of the bone intensifying method to strengthen bone,
The bone intensifying method includes absorbing protein material according to claim 1 with the amount of more than 5mg/ days.
5. a kind of method for preparing protein material according to claim 1,1)~3 it comprises the following steps)：
1) angiogenin and/or angiogenesis cellulose hydrolysate are prepared；
2) cystatin and/or cystatin hydrolysate are prepared；With
3) mixing according to it is above-mentioned 2) described in cystatin and/or cystatin hydrolysis production
Thing and according to it is above-mentioned 1) described in angiogenin and/or angiogenesis cellulose hydrolysate, with cause with angiogenin and/or
The mass ratio of angiogenesis cellulose hydrolysate is 0.003~0.6.
6. a kind of method for manufacturing protein material according to claim 1, it is included from breast and/or in the material from breast
Extraction includes angiogenin and/or angiogenesis cellulose hydrolysate and cystatin and/or half Guang of suppression
The fraction of serine protease protein hydrolysate, to cause the quality with angiogenin and/or angiogenesis cellulose hydrolysate
Than for 0.003~0.6 the step of.
7. according to the method for claim 6, its further comprise the angiogenin being included in the fraction and/or
The other step of cystatin enzymatic decomposition.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710552855.9A CN107446035A (en) | 2012-07-31 | 2012-07-31 | New protein material |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710552855.9A CN107446035A (en) | 2012-07-31 | 2012-07-31 | New protein material |
| CN201280074973.9A CN104507333A (en) | 2012-07-31 | 2012-07-31 | Novel protein material |
| PCT/JP2012/069391 WO2014020675A1 (en) | 2012-07-31 | 2012-07-31 | Novel protein material |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201280074973.9A Division CN104507333A (en) | 2012-07-31 | 2012-07-31 | Novel protein material |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107446035A true CN107446035A (en) | 2017-12-08 |
Family
ID=50027407
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710552855.9A Pending CN107446035A (en) | 2012-07-31 | 2012-07-31 | New protein material |
| CN201280074973.9A Pending CN104507333A (en) | 2012-07-31 | 2012-07-31 | Novel protein material |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201280074973.9A Pending CN104507333A (en) | 2012-07-31 | 2012-07-31 | Novel protein material |
Country Status (16)
| Country | Link |
|---|---|
| US (3) | US20150258184A1 (en) |
| EP (1) | EP2880994B1 (en) |
| JP (1) | JP6203723B2 (en) |
| KR (4) | KR20190062611A (en) |
| CN (2) | CN107446035A (en) |
| AU (1) | AU2012386758C1 (en) |
| BR (1) | BR112015002048A2 (en) |
| CA (1) | CA2879959C (en) |
| DK (1) | DK2880994T3 (en) |
| HK (1) | HK1207258A1 (en) |
| MX (1) | MX2015001343A (en) |
| MY (1) | MY190107A (en) |
| PH (1) | PH12015500048B1 (en) |
| SG (1) | SG11201500445SA (en) |
| TW (1) | TWI574696B (en) |
| WO (1) | WO2014020675A1 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP6941417B2 (en) * | 2016-02-04 | 2021-09-29 | 雪印メグミルク株式会社 | Cartilage function improver |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0362259A1 (en) * | 1987-05-22 | 1990-04-11 | Novo Nordisk As | Method of producing cystatin c or modifications hereof and dna-sequence for use when carrying out the method. |
| US20030206963A1 (en) * | 1999-03-30 | 2003-11-06 | Yukihiro Takada | Bone resorption suppressing agent |
| CN102099050A (en) * | 2008-05-14 | 2011-06-15 | 维多利亚农业服务控股公司 | Orally administrable dosage forms comprising angiogenin and uses thereof |
Family Cites Families (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4897464A (en) * | 1985-04-17 | 1990-01-30 | President And Fellows Of Harvard College | Purified protein having angiogenic activity and methods of preparation |
| JP3112637B2 (en) | 1994-09-30 | 2000-11-27 | 雪印乳業株式会社 | Bone strengthener |
| JP3929088B2 (en) | 1996-06-20 | 2007-06-13 | 雪印乳業株式会社 | Bone formation promoter and bone resorption inhibitor |
| JP4647750B2 (en) * | 2000-06-20 | 2011-03-09 | 雪印乳業株式会社 | Fraction containing high amount of milk basic cystatin and method for producing degradation product thereof |
| JP2001346519A (en) * | 2000-06-09 | 2001-12-18 | Snow Brand Milk Prod Co Ltd | Method for producing fraction containing high content of milk basic cystatin and decomposed product thereof |
| US6649590B2 (en) * | 2000-06-09 | 2003-11-18 | Snow Brand Milk Products Co., Ltd. | Method of producing fractions containing a high concentration of milk basic cystatin and decomposition products thereof |
| EP1602284A1 (en) * | 2000-06-09 | 2005-12-07 | Snow Brand Milk Products, Co., Ltd. | Method of producing fractions containing a high concentration of milk basic cystatin and decomposition products thereof |
| RU2519645C2 (en) * | 2008-05-14 | 2014-06-20 | Эгрикалчер Виктория Сервисиз Пти Лтд | Application of angiogenin or angiogenin agonists for treating diseases and disorders |
| NZ589311A (en) * | 2008-05-14 | 2012-08-31 | Agriculture Victoria Serv Pty | Angiogenin-enriched milk fractions prepared by methods involving heating the milk to over 70 degrees celsius for at least one minute |
| KR20100084454A (en) * | 2009-06-24 | 2010-07-26 | 원광대학교산학협력단 | Composition for reproduction of bone containing angiogenin |
-
2012
- 2012-07-31 KR KR1020197015142A patent/KR20190062611A/en not_active Application Discontinuation
- 2012-07-31 DK DK12882383.8T patent/DK2880994T3/en active
- 2012-07-31 EP EP12882383.8A patent/EP2880994B1/en active Active
- 2012-07-31 CN CN201710552855.9A patent/CN107446035A/en active Pending
- 2012-07-31 BR BR112015002048A patent/BR112015002048A2/en not_active Application Discontinuation
- 2012-07-31 MX MX2015001343A patent/MX2015001343A/en unknown
- 2012-07-31 CN CN201280074973.9A patent/CN104507333A/en active Pending
- 2012-07-31 US US14/418,276 patent/US20150258184A1/en not_active Abandoned
- 2012-07-31 JP JP2014527848A patent/JP6203723B2/en active Active
- 2012-07-31 KR KR1020187020460A patent/KR20180084159A/en active Application Filing
- 2012-07-31 WO PCT/JP2012/069391 patent/WO2014020675A1/en active Application Filing
- 2012-07-31 KR KR1020207002224A patent/KR102277774B1/en active IP Right Grant
- 2012-07-31 MY MYPI2015700262A patent/MY190107A/en unknown
- 2012-07-31 KR KR1020157004149A patent/KR20150036688A/en not_active Application Discontinuation
- 2012-07-31 AU AU2012386758A patent/AU2012386758C1/en active Active
- 2012-07-31 SG SG11201500445SA patent/SG11201500445SA/en unknown
- 2012-07-31 CA CA2879959A patent/CA2879959C/en active Active
-
2013
- 2013-07-25 TW TW102126683A patent/TWI574696B/en active
-
2015
- 2015-01-08 PH PH12015500048A patent/PH12015500048B1/en unknown
- 2015-08-13 HK HK15107833.9A patent/HK1207258A1/en unknown
-
2018
- 2018-11-29 US US16/204,155 patent/US20190091304A1/en not_active Abandoned
-
2021
- 2021-09-03 US US17/466,241 patent/US20210393751A1/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0362259A1 (en) * | 1987-05-22 | 1990-04-11 | Novo Nordisk As | Method of producing cystatin c or modifications hereof and dna-sequence for use when carrying out the method. |
| US20030206963A1 (en) * | 1999-03-30 | 2003-11-06 | Yukihiro Takada | Bone resorption suppressing agent |
| CN102099050A (en) * | 2008-05-14 | 2011-06-15 | 维多利亚农业服务控股公司 | Orally administrable dosage forms comprising angiogenin and uses thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CA2879959C (en) | 2017-03-14 |
| MY190107A (en) | 2022-03-29 |
| NZ704905A (en) | 2016-01-29 |
| TW201408318A (en) | 2014-03-01 |
| EP2880994A1 (en) | 2015-06-10 |
| EP2880994A4 (en) | 2016-01-20 |
| US20190091304A1 (en) | 2019-03-28 |
| KR20150036688A (en) | 2015-04-07 |
| KR20200011583A (en) | 2020-02-03 |
| BR112015002048A2 (en) | 2017-07-04 |
| PH12015500048A1 (en) | 2015-03-02 |
| KR20180084159A (en) | 2018-07-24 |
| US20150258184A1 (en) | 2015-09-17 |
| KR102277774B1 (en) | 2021-07-23 |
| KR20190062611A (en) | 2019-06-05 |
| AU2012386758B2 (en) | 2016-05-05 |
| WO2014020675A1 (en) | 2014-02-06 |
| MX2015001343A (en) | 2015-07-23 |
| CA2879959A1 (en) | 2014-02-06 |
| EP2880994B1 (en) | 2017-06-28 |
| SG11201500445SA (en) | 2015-04-29 |
| AU2012386758C1 (en) | 2017-03-30 |
| JPWO2014020675A1 (en) | 2016-07-11 |
| JP6203723B2 (en) | 2017-09-27 |
| US20210393751A1 (en) | 2021-12-23 |
| PH12015500048B1 (en) | 2015-03-02 |
| CN104507333A (en) | 2015-04-08 |
| AU2012386758A1 (en) | 2015-03-05 |
| DK2880994T3 (en) | 2017-10-02 |
| TWI574696B (en) | 2017-03-21 |
| HK1207258A1 (en) | 2016-01-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US9861687B2 (en) | Protein material | |
| TWI620509B (en) | Use of beverage and method of manufacturing same | |
| US20210393751A1 (en) | Novel protein material | |
| JP6357266B2 (en) | Protein material for bone disease prevention or treatment | |
| JP6357265B2 (en) | Protein material for bone disease prevention or treatment | |
| NZ704905B2 (en) | Protein material comprising angiogenin and cystatin | |
| NZ704911B2 (en) | Protein material comprising angiogenin and lactoperoxidase |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20171208 |
|
| RJ01 | Rejection of invention patent application after publication |